OPA1 and MICOS Regulate Mitochondrial Crista Dynamics And
Total Page：16
File Type：pdf, Size：1020Kb
Hu et al. Cell Death and Disease (2020) 11:940 https://doi.org/10.1038/s41419-020-03152-y Cell Death & Disease ARTICLE Open Access OPA1 and MICOS Regulate mitochondrial crista dynamics and formation Chao Hu1,LiShu1, Xiaoshuai Huang2, Jianglong Yu1, liuju Li2, Longlong Gong1, Meigui Yang1, Zhida Wu1, Zhi Gao1, Yungang Zhao3, Liangyi Chen2 and Zhiyin Song1 Abstract Mitochondrial cristae are the main site for oxidative phosphorylation, which is critical for cellular energy production. Upon different physiological or pathological stresses, mitochondrial cristae undergo remodeling to reprogram mitochondrial function. However, how mitochondrial cristae are formed, maintained, and remolded is still largely unknown due to the technical challenges of tracking mitochondrial crista dynamics in living cells. Here, using live-cell Hessian structured illumination microscopy combined with transmission electron microscopy, focused ion beam/ scanning electron microscopy, and three-dimensional tomographic reconstruction, we show, in living cells, that mitochondrial cristae are highly dynamic and undergo morphological changes, including elongation, shortening, fusion, division, and detachment from the mitochondrial inner boundary membrane (IBM). In addition, we find that OPA1, Yme1L, MICOS, and Sam50, along with the newly identified crista regulator ATAD3A, control mitochondrial crista dynamics. Furthermore, we discover two new types of mitochondrial crista in dysfunctional mitochondria, “cut- through crista” and “spherical crista”, which are formed due to incomplete mitochondrial fusion and dysfunction of the MICOS complex. Interestingly, cut-through crista can convert to “lamellar crista”. Overall, we provide a direct link between mitochondrial crista formation and mitochondrial crista dynamics. 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Introduction establish and maintain their unique structure to provide a Mitochondria contain outer and inner membranes. The large amount of surface area for chemical reactions to inner mitochondrial membrane is organized in two occur. Mitochondrial cristae have an orderly arrangement morphologically distinct regions: the inner boundary and contain respiratory chain supercomplexes, cyto- membrane (IBM) and the crista membrane (CM)1. The chrome c, and certain proteins that regulate mitochon- IBM contains translocases and certain proteins, such as drial oxidative phosphorylation and electron transfer3,4. OXA1 and Mia40, to shuttle proteins into the mito- Three-dimensional (3D) images show that mitochondrial chondrial matrix2. The CM is usually connected to IBM cristae are bag-like structures1,5. by the crista junction (CJ), which is a narrow constric- Mitochondrial cristae have long been considered tubu- tion3,4. Mitochondrial cristae harbor proteins that lar or lamellar invaginations formed by protrusion of the IBM into the mitochondrial matrix. OPA1, dimeric F1FO- ATP synthase, and MICOS (mitochondrial contact site Correspondence: Liangyi Chen ([email protected])or Zhiyin Song ([email protected]) and cristae organizing system) complex play key roles – 1Hubei Key Laboratory of Cell Homeostasis, Frontier Science Center for during this process6 8. OPA1 oligomerization narrows the Immunology and Metabolism, College of Life Sciences, Wuhan University, CJ and promotes ATP synthase dimerization9,10. The Wuhan, Hubei 430072, China 2State Key Laboratory of Membrane Biology, Beijing Key Laboratory of dimerization of ATP synthase is necessary to control Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking crista biogenesis and morphology, and it induces a posi- University, Beijing, China tive curvature that results in the formation of a crista Full list of author information is available at the end of the article 6 These authors contributed equally: Chao Hu, Li Shu sheet at the tip . The MICOS complex regulates the Edited by M. Campanella © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a linktotheCreativeCommons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Official journal of the Cell Death Differentiation Association Hu et al. Cell Death and Disease (2020) 11:940 Page 2 of 17 formation of CJs and mediates the contact between the study the dynamic changes in mitochondrial cristae in living OMM and IMM by interacting with the SAM (the sorting cells. We found that the membrane components of the and assembly machinery) complex, which is located on mitochondrial IBM and cristae membrane were not evenly the mitochondrial outer membrane and functions in the distributed within the mitochondria (Figs. S1A–S1C). assembly of β-barrel proteins into the mitochondrial outer Therefore, communication and material exchange between membrane11,12. The MICOS complex consists of at least the mitochondrial cristae and IBM should occur to maintain 7 subunits in mammals: Mic60, Mic27, Mic26, Mic25, mitochondrial membrane homeostasis. In living cells, mito- Mic19, Mic13, and Mic103. Recently, Harner et al. chondrial cristae were mainly tubular or lamellar, highly hypothesized that mitochondrial fusion and the MICOS dynamic, and organized, with remarkable changes in length complex contribute to the formation of mitochondrial and position within one second (Fig. 1A–C and Supple- lamellar cristae7. mentary Video 1). The mitochondrial cristae constantly Under normal conditions, normal tubular or lamellar altered their lengths by elongation or shortening, detached mitochondrial cristae have an orderly arrangement. from the IBM, or contacted and fused with the IBM (Fig. However, under different physiological and pathological 1A–C and S1D), suggesting that mitochondrial cristae are conditions, the number, size, shape, and distribution of dynamically altered in length and constantly contact and mitochondrial cristae are affected to a very different communicate with the IBM. Moreover, two mitochondrial extent1,3,4,9,13. For example, in aged animals, mitochon- cristae could fuse into one (Fig. 1D), and one crista could drial cristae are enlarged, and ~25% of mitochondria divide into two (Fig. 1E). Occasionally, mitochondrial cristae contain large central voids that lack CJs14,15. However, the contacted each other and fused to form a crista network; mechanism and functions of mitochondrial crista remo- however, this contact was subsequently separated (Fig. 1F), deling are largely unclear. suggesting that mitochondrial cristae contact each other via a Mitochondria continuously fuse and divide to maintain “kiss-and-run” mode. We also measured the mitochondrial normal mitochondrial morphology and functions. Mitofusins crista movement rate and found that mitochondrial IBMs (Mfn1 and Mfn2) and OPA1 mediate outer and inner had a substantial overlap, but a few mitochondrial cristae did mitochondrial membrane fusion, respectively16,17. Addition- not, as seen by comparing the 0-sec and 0.1-sec images (Fig. ally, Drp1 is a key regulator of mitochondrial fission18,19. 1G). However, most mitochondrial cristae showed no over- Theoretically, similar to mitochondria, mitochondrial cristae lap between the 0-sec and 2-sec images (Fig. 1H). Addi- should be highly dynamic. However, due to the limitation of tionally, mitochondrial cristae can undergo elongation, technology in detecting mitochondrial cristae in living cells, shortening, detachment, fusion, and fission in one or a few mitochondrial crista dynamics are not fully understood. seconds (Fig. 1A–F). These data suggest that mitochondrial For more than half a century, mitochondrial cristae have cristae are highly dynamic and change within a few seconds. only been analyzed by transmission electron microscopy Mitochondria can contact each other by a “kiss-and-run” (TEM), which cannot be applied to living cells. Recently, mode28. We then investigated whether mitochondrial cristae the development of super-resolution microscopy tech- are involved in mitochondrion–mitochondrion contact by nology has provided an excellent tool for visualizing Hessian-SIM. During mitochondrion–mitochondrion con- – mitochondria or mitochondrial cristae in living cells20 26. tact (kiss-and-run fusion), direct mitochondrial crista–crista We previously established a super-resolution microscopy contact was observed (Fig. 2A). Moreover, during mito- technology called structured illumination microscopy chondrial fusion, mitochondrial cristae contact and fuse with (Hessian-SIM), which enables fast, long-term, and super- each other (Fig. 2B). Our data are consistent with a previous resolution imaging of mitochondrial cristae27. Here, we report indicating that the cristae of adjacent mitochondria used Hessian-SIM combined with TEM, focused ion are coordinated at inter-mitochondrial junctions in cardio- beam/scanning electron microscopy (FIB-SEM), and 3D myocytes29.Overall,thesefindingssuggestthatmitochon-